• Title/Summary/Keyword: C2C12 cell

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Antimutagenic and Anticancer Effects of Leaf Mustard and Leaf Mustard Kimchi

  • Kim, Yong-Taek;Kim, Boh-Kyung;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.12 no.2
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    • pp.84-88
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    • 2007
  • In this study, we investigated antimutagenic and anticancer activities of leaf mustard (LM, Brassica juncea) and leaf mustard kimchi (LMK) during their fermentation period. Methanol extracts were prepared from raw mustard, brined leaf mustard in 10% Gueun salt solution for 2 hrs, leaf mustard fermented at 15$^{\circ}C$ for 5 days after brined in 10% Guenun salt solution for 2 hrs (Fr-LM), fresh leaf mustard kimchi (Fresh-LMK) and optimally ripened leaf mustard kimchi fermented at 5$^{\circ}C$ for 30 days (OR-LMK). OR-LMK showed the strongest inhibitory activities against the mutagenicities induced by aflatoxin B1 in Salmonella Typhimurium TA100. LMs and LMKs inhibited the survival or growth of AGS human gastric adenocarcinoma cells and HT-29 human colon carcinoma cells in MTT assay and growth inhibition test. Among the extracts, OR-LMK and FR-LM exhibited strong antiproliferative effect against cancer cells, especially HT-29 cells. DAPI staining assay showed that OR-LMK induced apoptosis cell death of HT-29 cells in a dose-dependent manner. These results suggest that leaf mustards and leaf mustard kimchi have chemopreventive activities.

PKC-Independent Stimulation of Cardiac $Na^+/Ca^{2+}$ Exchanger by Staurosporine

  • Kang, Tong-Mook
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.5
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    • pp.259-265
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    • 2008
  • $[Ca^{2+}]_i$ transients by reverse mode of cardiac $Na^+/Ca^{2+}$ exchanger (NCX1) were recorded in fura-2 loaded BHK cells with stable expression of NCX1. Repeated stimulation of reverse NCX1 produced a long-lasting decrease of $Ca^{2+}$ transients ('rundown'). Rundown of NCX1 was independent of membrane $PIP_2$ depletion. Although the activation of protein kinase C (PKC) was observed during the $Ca^{2+}$ transients, neither a selective PKC inhibitor (calphostin C) nor a PKC activator (PMA) changed the degrees of rundown. By comparison, a non-specific PKC inhibitor, staurosporine (STS), reversed rundown in a dose-dependent and reversible manner. The action of STS was unaffected by pretreatment of the cells with calphostin C, PMA, or forskolin. Taken together, the results suggest that the stimulation of reverse NCX1 by STS is independent of PKC and/or PKA inhibition.

The anti-inflammation effects of A.C.C. extracts on the LPS-induced Raw 264.7 cell (LPS로 유도한 Raw 264.7 세포에서 A.C.C. 추출물의 항염증 효과)

  • Ryu, Jin-Hyeob;An, Ju-Hee;Woo, Yong-Kyu;Cho, Hyun-Jeong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.12
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    • pp.503-511
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    • 2017
  • This study was conducted to evaluate the anti-inflammatory activity and clinical efficacy of a sample (A.C.C. extracts) obtained by distillation extraction of 14 herbal medicines including Phellodendron bark. To confirm this, the amount of nitric oxide (NO) produced by the cells in RAW 264.7 cells stimulated by lipopolysaccharide (LPS) and the changes in the production of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$, interleukin(IL)-$1{\beta}$ and IL-6 were determined. The results showed that A.C.C. extracts strongly inhibited the production of NO and inflammatory cytokines increased by LPS without cytotoxicity. In addition, A.C.C. extracts showed strong bacterial reduction rates of 99.9% in Pseudomonas aeruginosa, Staphylococcus aureus, MRSA (Methicillin-resistant Staphylococcus aureus), Candida albicans and Streptococcus mutans. These findings indicate that A.C.C. extracts are effective ingredients with a strong antimicrobial effect together with an anti-inflammatory effect. In addition, when A.C.C. extracts were applied to infants and toddlers who were suffering from diaper rash, itching, and perspiration symptoms, symptoms of rash, atopy, rash, itching, and heat rash were improved. After the lapse of time, it was visually confirmed that it was considerably relaxed. These findings confirm that A.C.C. extracts comprise a clinically effective anti-inflammatory and anti-bacterial agent that alleviates symptoms such as diaper rash and fever and may therefore be an effective alternative to inflammatory diseases.

Influence of Water Temperature and Salinity on Egg Development of Flatfish, Limanda herzensteini (참가자미, Limanda herzensteini의 난발생에 미치는 수온과 염분의 영향)

  • 이정용;김완기;장영진
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.357-362
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    • 1997
  • In order to obtain the basic information for seedling production of flatfish, Limanda herzensteini, the influence of water temperature and salinity on egg development was investigated. The desirable water temperature for egg hatching was9~$15^\circC$. The time of egg development was shorter with higher water temperature. The relationships between the water temperature (T:$^\circC$) and the required time (t:hour) from egg to each development stage were given as follows ; 8-cell : 1/t=0.0284T-0.0554 (r=0.9999) Morula : 1/t=0.0137T-0.0527 (r=0.9998) Kupffer's vesicle : 1/t=0.0035T-0.0133 (r=0.9762) Hatching : 1/t=0.0012T-0.0007 (r=0.9981) Biological mimimum temperature for the egg development was estimated to the be $2.6^\circC$ in average. The salinity which showed over 50% survival rate from fertilized egg to hatching was 35~$38\textperthousand$.

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Studies on Cryopreservation of 8-Cell Mouse Embryos (생쥐 8세포기 수정란의 동결보존에 관한 연구)

  • 최미후;이규승;박칭식;서길웅
    • Korean Journal of Animal Reproduction
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    • v.11 no.3
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    • pp.155-160
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    • 1987
  • This study was carried out to determine the effects of cryoprotectants, freezing and thawing rates on the survival of 8-cell mouse embryos. The female ICR mice were induced to superovulated by intraperitoneal injections of 5 i.u. PMSG and 5 i.u. HCG given 48h apart and then were paired with males of the same strain. They were killed and embryos were flushed from the oviducts and uteri on 3 days after injection of HCG. Embryos were flushed with modified Dulbecco's phosphate buffered saline and equilibrated with 1.5M-dimethyl sulfoxide (DMSO) or 1.5M-glycerol by 3-step procedure. The freezing rates of the embryos were 1$^{\circ}C$/min from room temperature to -5$^{\circ}C$ and the embryos were seeded at -5$^{\circ}C$. After being held for 3 min at the seeding temperature, the rates were 0.3$^{\circ}C$/min from -5$^{\circ}C$ to -35$^{\circ}C$. From -35$^{\circ}C$ to -7$0^{\circ}C$, the rates were divided into 0.1$^{\circ}C$/min, 1$^{\circ}C$/min and 1$0^{\circ}C$/min, respectively. After being held for 5 min at -7$0^{\circ}C$, the embryos were plunged directly into liquid nitrogen. The embryos were thawed at 4$^{\circ}C$/min and 12$^{\circ}C$/min from -196$^{\circ}C$ to 37$^{\circ}C$, and for 2 min in 37$^{\circ}C$ water bath, respectively. The average number of ovulation points and embryos recovered were 42.7 and 34 appearing 79.5% recovery rate. Eight cell embryos in the embryos recovered were 26.3. The survival rates of embryos according to the freezing rates in the presence of 1.5M-DMSO were 73.5~80.6% at 0.1$^{\circ}C$/min, 75.0~79.5% at 1$^{\circ}C$/min and 52.8~54.7% at 1$0^{\circ}C$/min, but in the presence of 1.5M-glycerol were 62.9~67.6% at 0.1$^{\circ}C$/min, 61.4~68.3% at 1$^{\circ}C$/min and 25.5~30.2% at 1$0^{\circ}C$/min. The survival rates of embryos were not affected by the thawing rates.

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The Adenylyl Cyclase Activator Forskolin Increases Influenza Virus Propagation in MDCK Cells by Regulating ERK1/2 Activity

  • Sang-Yeon Lee;Jisun Lee;Hye-Lim Park;Yong-Wook Park;Hun Kim;Jae-Hwan Nam
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1576-1586
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    • 2023
  • Vaccination is the most effective method for preventing the spread of the influenza virus. Cell-based influenza vaccines have been developed to overcome the disadvantages of egg-based vaccines and their production efficiency has been previously discussed. In this study, we investigated whether treatment with forskolin (FSK), an adenylyl cyclase activator, affected the output of a cell-based influenza vaccine. We found that FSK increased the propagation of three influenza virus subtypes (A/H1N1/California/4/09, A/H3N2/Mississippi/1/85, and B/Shandong/7/97) in Madin-Darby canine kidney (MDCK) cells. Interestingly, FSK suppressed the growth of MDCK cells. This effect could be a result of protein kinase A (PKA)-Src axis activation, which downregulates extracellular signal-regulated kinase (ERK)1/2 activity and delays cell cycle progression from G1 to S. This delay in cell growth might benefit the binding and entry of the influenza virus in the early stages of viral replication. In contrast, FSK dramatically upregulated ERK1/2 activity via the cAMP-PKA-Raf-1 axis at a late stage of viral replication. Thus, increased ERK1/2 activity might contribute to increased viral ribonucleoprotein export and influenza virus propagation. The increase in viral titer induced by FSK could be explained by the action of cAMP in assisting the entry and binding of the influenza virus. Therefore, FSK addition to cell culture systems could help increase the production efficiency of cell-based vaccines against the influenza virus.

Development of a New Air Cooling System Utilizing the Stirling Engine for Preventing Solar Cell from Overheating (태양광 모듈의 과열 방지용 공랭형 스털링기관 냉각 시스템 개발)

  • Kim, Hyoungeun;Park, Chanwoo;Chu, Jinkyung;Keum, Dongyeop;Park, Silro;Kim, Jeongmin;Kim, Daejin
    • Transactions of the KSME C: Technology and Education
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    • v.2 no.1
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    • pp.57-63
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    • 2014
  • In this paper new air-cooling system utilizing Stirling engine was proposed for improving efficiency in solar photovoltaic power generation. The solar cell plate was equipped with semi-circular channel for air flow on the backside. Beta-type Stirling engine was installed on the plate and its flywheel was connected to a motor fan by a transmission belt. A forced convective air flow for heat radiation was generated by the operation of the self-starting Stirling engine. The performance tests for power generation of solar cell with or without the proposed air-cooling system were conducted under halogen lamp. From the experimental results, it was found that decline in output voltage of the solar cell with proposed cooling system was 25% less than that of the solar cell without cooling system.

Overexpression of CXCL2 inhibits cell proliferation and promotes apoptosis in hepatocellular carcinoma

  • Ding, Jun;Xu, Kangdi;Zhang, Jie;Lin, Bingyi;Wang, Yubo;Yin, Shengyong;Xie, Haiyang;Zhou, Lin;Zheng, Shusen
    • BMB Reports
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    • v.51 no.12
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    • pp.630-635
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    • 2018
  • C-X-C motif chemokine ligand 2 (CXCL2) is a small secreted protein that exhibits a structure similar to the proangiogenic subgroup of the CXC chemokine family. Recently, accumulating evidence suggests that chemokines play a pivotal role in cancer progression and carcinogenesis. We examined the expression levels of 7 types of $ELR^+$ CXCLs messenger RNA (mRNA) in 264 clinical samples. We found that CXCL2 expression was stably down-regulated in 94% of hepatocellular carcinoma (HCC) specimens compared with paired adjacent normal liver tissues and some HCC cell lines. Moreover, CXCL2 overexpression profoundly attenuated HCC cell proliferation and growth and induced apoptosis in vitro. In animal studies, we found that overexpressing CXCL2 by lentivirus also apparently inhibited the size and weight of subcutaneous tumours in nude mice. Furthermore, we demonstrated that CXCL2 induced HCC cell apoptosis via both nuclear and mitochondrial apoptosis pathways. Our results indicate that CXCL2 negatively regulates the cell cycle in HCC cells via the ERK1/2 signalling pathway. These results provide new insights into HCC and may ultimately lead to the discovery of innovative therapeutic approaches of HCC.

MiR-141-3p regulates myogenic differentiation in C2C12 myoblasts via CFL2-YAP-mediated mechanotransduction

  • Nguyen, Mai Thi;Lee, Wan
    • BMB Reports
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    • v.55 no.2
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    • pp.104-109
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    • 2022
  • Skeletal myogenesis is essential to keep muscle mass and integrity, and impaired myogenesis is closely related to the etiology of muscle wasting. Recently, miR-141-3p has been shown to be induced under various conditions associated with muscle wasting, such as aging, oxidative stress, and mitochondrial dysfunction. However, the functional significance and mechanism of miR-141-3p in myogenic differentiation have not been explored to date. In this study, we investigated the roles of miR-141-3p on CFL2 expression, proliferation, and myogenic differentiation in C2C12 myoblasts. MiR-141-3p appeared to target the 3'UTR of CFL2 directly and suppressed the expression of CFL2, an essential factor for actin filament (F-actin) dynamics. Transfection of miR-141-3p mimic in myoblasts increased F-actin formation and augmented nuclear Yes-associated protein (YAP), a key component of mechanotransduction. Furthermore, miR-141-3p mimic increased myoblast proliferation and promoted cell cycle progression throughout the S and G2/M phases. Consequently, miR-141-3p mimic led to significant suppressions of myogenic factors expression, such as MyoD, MyoG, and MyHC, and hindered the myogenic differentiation of myoblasts. Thus, this study reveals the crucial role of miR-141-3p in myogenic differentiation via CFL2-YAP-mediated mechanotransduction and provides implications of miRNA-mediated myogenic regulation in skeletal muscle homeostasis.

The effects of Allomyrina dichotoma larval extract on palmitate-induced insulin resistance in skeletal muscle cells (장수풍뎅이 유충 추출물이 고지방산 처리 골격근세포의 인슐린 저항성에 미치는 영향)

  • Kim, Kyong;Sim, Mi-Seong;Kwak, Min-Kyu;Jang, Se-Eun;Oh, Yoon Sin
    • Journal of Nutrition and Health
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    • v.55 no.4
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    • pp.462-475
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    • 2022
  • Purpose: Allomyrina dichotoma larvae are one of the approved edible insects with nutritional value and various functional and medicinal properties. Previously we have demonstrated that the Allomyrina dichotoma larval extract (ADLE) ameliorates hepatic insulin resistance in high-fat diet (HFD)-induced diabetic mice through the activation of adenosine monophosphate-activated protein kinase (AMPK). This study investigated the effects of ADLE on insulin resistance in the skeletal muscle and explored mechanisms for enhancing the glucose uptake in palmitate (PAL)-treated C2C12 myotubes. Methods: To induce insulin resistance, the differentiated C2C12 myotubes were treated with PAL (0.5 mM) for 24 hours, and then treated with a 0.5 mg/ml concentration of ADLE, and the resultant effects were measured. The expression levels of glucose transporter-4 (GLUT4), AMPK, and the mitochondrial metabolism-related proteins were analyzed by western blotting. The mRNA expression levels of lipogenesis- related genes were determined by quantitative reverse-transcriptase PCR. Results: The exposure of C2C12 myotubes to 0.5 mg/ml of ADLE increased cell viability significantly compared to PAL-treated cells. ADLE upregulated the protein expression of GLUT4 and enhanced glucose uptake in the PAL-treated cells. ADLE increased the phosphorylated AMPK in both the PAL-treated C2C12 myotubes and HFD-treated skeletal muscle. The reduced expression levels of peroxisome-proliferator-activated receptor gamma co-activator-1 alpha (PGC1α) and uncoupling protein 3 (UCP3) due to the PAL and HFD treatment were reversed by the ADLE treatment. The citrate synthase activity was also significantly increased with the PAL and ADLE co-treatment. Moreover, the mRNA and protein expressions of fatty acid synthesis-related factors were reduced in the PAL and HFD-treated muscle cells, and this effect was significantly attenuated by the ADLE treatment. Conclusion: ADLE activates AMPK, which in turn induces mitochondrial metabolism and reduces fatty acid synthesis in C2C12 myotubes. Therefore, ADLE could be useful for preventing or treating insulin resistance of skeletal muscles in diabetes.